Effects of PI3K inhibitor, Akt inhibitor or AdPTEN on the ERβ agonist-induced vasodepressive responses. Time-course of the changes in MSAP and total power density of vasomotor components (0–0.8 Hz) of SAP spectrum in anaesthetized rats that received microinjection bilaterally into the RVLM (at time 0) of aCSF, or DPN (2 pmol) given together with LY294002 (LY, 5 pmol), Akt inhibitor (Akt I, 250 pmol) or 5 % DMSO (A), or with additional treatment with AdPTEN (5 × 108 pfu) or AdGFP (5 × 108 pfu), administered into the bilateral RVLM 7 days before DPN (2 pmol) microinjection (B). (C) Representative Western blots (insets) or densitometric analysis of the amount of PTEN protein relative to basal PTEN protein levels (control), detected from the RVLM 3, 7 or 14 days after animals received microinjection bilaterally into the RVLM of AdPTEN (5 × 108 pfu) or AdGFP (5 × 108 pfu). (D) Representative photomicrographs showing the distribution of PTEN-immunoreactivity (arrows) in RVLM on 7 days after animals received bilateral microinjection into the RVLM of aCSF (control), AdPTEN (5 × 108 pfu) or AdGFP (5 × 108 pfu). Calibration bar: 100 μm in low magnification and 50 μm in high magnification of inset. Values are presented as the mean ± SEM; n = 6-7 animals per experimental group. *P < 0.05 versus aCSF or control group and #P < 0.05 versus DPN + DMSO or AdGFP + DPN group in the post hoc Scheffé multiple-range test. NA, nucleus ambiguus; RVLM, rostral ventrolateral medulla.